The need for the development of new strategies for treatment of bladder cancer (BC) has experienced a palpable increase in the last few years. While several options are currently under investigation, many if not all remain suboptimal. This is critical as it is estimated that in 2015 there were ~450,000 new cases and over 115,000 deaths from BC worldwide. In the US alone there were ~74,000 new cases and over 16,000 deaths from BC. This makes BC the 6th most prevalent and 9th deadliest cancer worldwide. While various treatment options currently exist, a number of issues have been reported including the high cost of treatment, patient discomfort, complications, incomplete treatment, procedural invasiveness and poor efficacy among others. With a 5yr survival rate for BC patients ranging from 6 to 65% yielding a reported ~115,000 deaths in 2015 alone (far above prostate or breast cancer), there is an ever growing urgency and compelling need for the development of new strategies and devices to treat BC. The intent of this project is to develop a new cystoscopic based thermo-molecular treatment strategy and catheter system (FrostBite-BC) for the treatment of BC employing a minimally invasive trans-urethral outpatient procedure. The prototype unit and preliminary data collected to date strongly support the potential of this approach. The major goals of this Phase 1 proposal are the following: (1) Develop the FrostBite-BC catheter system and bench test it using non tissue engineering models. (2) Test this new treatment protocol in a homogeneous and heterogeneous cell population BC tissue engineered model (TEM). (3) Test the ability of an adjunctive nutraceutical compound, Vitamin D3, to sensitize BC cells to freeze injury (cryosensitization) thereby making cryoablation more effective while protecting normal cells and (4) conduct a pilot in vivo study to validate development. Phase 2 will include (1) optimizing the catheter, device and procedure, (2) conducting pre-clinical in vivo studies and (3) determining the potential role of FrostBite-BC launching a whole body immune response against metastatic disease in vivo. The successful completion of this project will lead to a new minimally invasive cystoscopic compatible trans-uretheral approach for the treatment of bladder and other urogenital cancers. Once completed, this project will provide for an effective, minimally invasive treatment for patients afflicted with BC as well as other urologic based cancers resulting in improved patient survival, quality of life, and mitigating risk factors for cancer diagnosis.